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        <title>Choosing a Cycling Regime</title>
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        <h1 align="center">Choosing a Cycling Regime </h1>
		<p>Despite the relatively simple principle of a cycling regime, there is 
		there are very few general guidelines as to how to choose an optimal 
		cycling regime. this is due in large part to universally accepted 
		guidelines </p>
		<p>One of the most prominent examples is the common almost inveterate 
		practice of including a primer annealing step somewhere below 60° C. 
		indeed, there is little if any justification for the necessity of such a 
		step and indeed search of early PCR literature provides no justification 
		for using such a low annealing temperatures. the most likely explanation 
		is that the cost of primer synthesis were high enough to justify 
		limiting primer size to the minimum to achieve specificity, which is 
		typically in the range of 18-20 bases. the melting temperatures of such 
		small primers were such that it was necessary to anneal at temperatures 
		below 60°C. however, it was a concern that such a low temperature would 
		limit enzyme activity and thus was borne the 72°C elongation step.</p>
		<p>An alternative approach is to simply increase the annealing 
		temperature to a level where loss in enzyme activity is much less of a 
		concern, allowing the elongation step to be eliminated. for reasons 
		discussed in the primer design section, for QuantiTect enzyme 
		formulation an annealing and elongation a standardized to&nbsp;65°C. </p>
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